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首页> 外文期刊>Gene expression >Determination of relative Notch1 and γ-secretase-related gene expression in puromycin-treated microdissected rat kidneys
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Determination of relative Notch1 and γ-secretase-related gene expression in puromycin-treated microdissected rat kidneys

机译：嘌呤霉素处理的显微解剖大鼠肾脏中Notch1和γ-分泌酶相关基因的相对表达的测定

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摘要

Notch signaling pathways are involved in the regulation of cell differentiation and are highly conserved across species. Notch ligand binding leads to γ-secretase-mediated proteolytic cleavage of the Notch receptor releasing the Notch intracellular domain, resulting in its subsequent translocation into the nucleus and gene expression regulation. To investigate the level of expression of Notch signaling pathway components in microanatomic regions following renal injury, kidneys from untreated, vehicle control, and puromycin aminonucleoside (PA, 150 mg/kg)-treated rats were evaluated. Frozen tissue sections from rats were microdissected using laser capture microdissection (LCM) to obtain glomeruli, cortical (proximal) tubules, and collecting ducts, and relative gene expression levels of Presenilin1, Notch1, and Hes1 were determined. In untreated rats, the Notch1 expression in glomeruli was higher than in the proximal tubules and similar to that in collecting ducts, whereas Presenilin1 and Hes1 expressions were highest in the collecting ducts, followed by cortical tubules and glomeruli. Following PA-induced renal injury, Hes1 gene expression increased significantly in the glomeruli and tubules compared to the collecting ducts where no injury was observed microscopically. Although these data present some evidence of change in Notch signaling related to injury, the expression of Presenilin1, Notch1, and Hes1 in the microanatomic regions of the kidney following PA treatment were not significantly different when compared to controls. These results demonstrate that there are differences in Notch-related gene expression in the different microanatomic regions of the kidneys in rats and suggest a minimal role for Notch in renal injury induced by PA. In addition, this work shows that LCM coupled with the RT-PCR can be used to determine the relative differences in target gene expression within regions of a complex organ.
机译：Notch信号通路参与细胞分化的调控，并且在物种间高度保守。 Notch配体结合导致Notch受体的γ-分泌酶介导的蛋白水解裂解，释放Notch细胞内结构域，导致其随后易位到核中并调节基因表达。为了研究Notch信号通路成分在肾脏损伤后的微解剖区域中的表达水平，评估了未处理，媒介物对照和嘌呤霉素氨基核苷（PA，150 mg / kg）处理的大鼠的肾脏。使用激光捕获显微切割术（LCM）对大鼠的冷冻组织切片进行显微解剖，以获得肾小球，皮层（近端）小管和收集管，并测定Presenilin1，Notch1和Hes1的相对基因表达水平。在未经治疗的大鼠中，肾小球中的Notch1表达高于近端小管，与收集管中的相似，而Presenilin1和Hes1的表达在收集管中最高，其次是皮质小管和肾小球。 PA诱发的肾损伤后，与未在显微镜下观察到损伤的收集管相比，肾小球和肾小管中Hes1基因的表达显着增加。尽管这些数据提供了一些与损伤相关的Notch信号变化的证据，但是与对照组相比，PA治疗后肾脏的微解剖区域中Presenilin1，Notch1和Hes1的表达没有显着差异。这些结果表明，在大鼠肾脏的不同微解剖区域中，Notch相关基因的表达存在差异，提示Notch在PA诱发的肾损伤中的作用很小。此外，这项工作表明，LCM与RT-PCR结合可用于确定复杂器官区域内靶基因表达的相对差异。

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